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Biomedical subjects

Y Brun

Publications and source records attributed to Y Brun.

At least 19 recordsLinked to original sources

Synthetic DNA probes for detection of genes for enterotoxins A, B, C, D, E and for TSST-1 in staphylococcal strains.

A dot blot hybridization technique with oligonucleotide probes was developed for the specific detection of the TSST-1 gene and the staphylococcal enterotoxin (SE) genes A, B, C, D and E. For each toxin gene a probe sequence was chosen from the previously determined sequence. A total of 145 staphylococcal strains (133 Staphylococcus aureus and 12 coagulase-negative staphylococci (CNS) were studied by this genotypic method and by two phenotypic assays (gel immunodiffusion and ELISA). An excellent correlation (96%) was observed between the genotypic and phenotypic assays. DNA from two CNS strains hybridized with a probe without detection of the corresponding toxin (SEB for one strain and SEC for the other strain). One Staph. aureus strain was shown to be an SEC producer, but was not detected by the corresponding probe. Gene probe and immunological assays seem to be complementary methods for studies of staphylococcal strains producing (or potentially producing) TSST-1 or enterotoxins.

Bacterial Toxins

Evaluation of a blood culture medium BHI-S-Lysis for BCB Release System Roche.

The new blood culture medium BHI-S Lysis for the BCB Release System (F. Hoffmann-La Roche AG., Basel, Switzerland), which lyses blood cells, was compared with the Hémoline performance Duo (BioMérieux, Marcy l'Etoile, France) including a diphasic medium and a modified Wilkins Chalgren broth. A total of 1998 sets were each inoculated with 7 ml of blood. In the laboratory, agar-coated paddles were attached to the BHI-S-Lysis. Overall, there was a statistically significant difference between the BHI-S-Lysis and the diphasic medium, or Wilkins Chalgren broth, in the recovery of clinically important microorganisms. The yield of pathogens with the BHI-S-Lysis, diphasic medium and Wilkins Chalgren broth was 115, 68 and 58, respectively. Staphylococci were detected by the BHI-S-Lysis significantly more frequently. Out of 50 bacteraemias, 12 were diagnosed with the BHI-S-Lysis only, three with the diphasic medium only, and three with the Wilkins Chalgren broth only. Contaminant isolates occurred with significantly greater frequency in the BHI-S-Lysis than in the two BioMérieux systems, in particular with coagulase-negative staphylococci. No speed advantage was present for all groups of micro-organisms and blood culture media. This good performance of the BHI-S-Lysis medium in detecting a larger number of pathogens should be confirmed by further evaluations.

Bacteremia

Difficulties in identifying Klebsiella strains of clinical origin.

Two hundred and four strains of Gram-negative bacteria of clinical origin, initially identified as Klebsiella using the API 20 E system, and 10 reference strains were further analysed with the API 20 EC test system and the API 50 CH, API 50 AO, API 50 AA assimilation systems. Four clusters corresponding to the species Klebsiella pneumoniae subsp. pneumoniae, K. oxytoca, K. planticola, and K. terrigena were formed after numerical analysis of 155 selected tests and the 26 most discriminating tests were determined. A comparison was made between conventional identification using the API 20 E system and the results of the numerical analysis. The conventional method resulted in incorrect identification of 13% of the strains tested, especially for the new species: K. planticola and K. terrigena. After numerical analysis, 17 out of 204 strains (8.3%) of clinical origin were identified as K. planticola. Only 1 strain of clinical origin was identified as K. terrigena, and 1 strain as K. ornithinolytica.

Humans

Characterization of clinically significant isolates of Staphylococcus epidermidis from patients with cerebrospinal fluid shunt infections.

Biotyping, slime production, antibiograms, extrachromosomal DNA banding and total DNA restriction analysis were used to characterize Staphylococcus epidermidis strains causing cerebrospinal fluid shunt infections in 11 patients. Infections considered to be community acquired and those acquired in the first 2 weeks of hospital admission were due to oxacillin-susceptible isolates. Multiply resistant strains were isolated from patients who were in hospital for more than 1 month before tube implantation. Slime was detected in staphylococci for 54% of cases, but its expression varied. Strains from different patients could be differentiated from one another by the extrachromosomal DNA bandings and total DNA restriction patterns, but isolates from the same patient were usually similar. During the period of external drainage, epidemiological markers were useful in differentiating persistence of infection from contamination or re-infection by a new strain.

Adult

Molecular epidemiology of Staphylococcus haemolyticus strains isolated in an Albanian hospital.

A recent outbreak of erythroderma in young children in an Albanian hospital was investigated. The etiology was not established, but Staphylococcus haemolyticus was frequently isolated from the affected children and from staff working in the same unit. Possible relationships among the isolates were investigated by using classical techniques (biotype, antimicrobial susceptibility, and extrachromosomal DNA pattern) and by restriction endonuclease analysis (REA) of total DNA. Control isolates of proven pathogenicity from hospitalized patients in Lyon, France were subjected to the same procedures. Distinct REA patterns were obtained after digestion with two enzymes in 7 of 10 isolates from five affected children. Six distinct patterns were observed in nine isolates from six staff members; two REA patterns from patient isolates and two from staff members were identical, and these were distinguishable by the other markers examined. Only two different REA patterns were found in the pathogenic control isolates despite the use of a third additional enzyme. Again, the isolates with the same REA patterns could be distinguished by their plasmid profile or antimicrobial resistance profile. REA of total DNA used in combination with other markers indicated that the Albanian isolates differed considerably, whereas the French pathogenic isolates showed little variability.

Albania

[In vitro antibacterial activity of a new fluoroquinolone, temafloxacin, against hospital isolates. Results of a multicenter study].

Minimal inhibitory concentrations (MICs) of temafloxacin (TMF) was determined by agar dilution for 2,510 bacterial strains isolated in 1989 in 9 university hospitals. Activity of TMF against nalidixic acid (NAL) susceptible (S) Enterobacteriaceae was close to that of other fluoroquinolones (FQ) (mode MIC: 0.06 micrograms/l); like for other FQ, this activity was reduced against NAL intermediate (mode 1) and resistant (R) (mode 4) Enterobacteriaceae. MICs of TMF against P. aeruginosa were between 0.12 and 128 (mode 0.5-1). TMF had also a good activity against NAL S A. baumannii (mode MIC: 0.06-0.12) but this activity is reduced against NAL R Acinetobacter (mode MIC: 16). TMF was highly active against Haemophilus mode MIC: less than or equal to 0.008), Gonococci (mode MIC: 0.008-0.032), Meningococci (mode MIC: 0.08) and B. catarrhalis (mode MIC: 0.016). TMF showed better activity to other fluoroquinolones against methicillin susceptible Staphylococci (mode MIC: 0.06); the resistant strains (mode MIC: 8) are usually methicillin resistant. Comparatively to the currently available FQ, TMF is so more effective against Enterococci (mode MIC: 1), Streptococci (mode MIC: 0.5-1) and Pneumococci (mode MIC: 0.5). Finally, for the anaerobic bacteria, TMF is more active against C. perfringens (mode MIC: 0.5) than against B. fragilis (mode MIC: 2).

4-Quinolones

International collaborative evaluation of the ATB 32 staph gallery for identification of the Staphylococcus species.

This international collaborative study evaluates a new system (ATB 32 Staph) for the identification of staphylococci taking into account the new novobiocin-sensitive and -resistant species reported. This study involved eight laboratories and 792 strains were tested. The reproducibility obtained for the cumulative results of the inter- and intra-laboratory tests was more than 90%. For 713 strains relevant of a species 95.5% were correctly identified by the system. Eight strains (1.2%) were misidentified and 24 strains (3.3%) were not identified. For 79 strains initially considered as not-classified, 62% were identified at the species level by the new system. The newer ATB 32 Staph gallery is a performant and useful method for routine identification of the currently described staphylococci species from clinical and animal origin.

Animals

Effectiveness of simple measures to control an outbreak of nosocomial methicillin-resistant Staphylococcus aureus infections in an intensive care unit.

Between June 1985 and March 1986, 14 cases of severe nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infection, including septicemia, were observed in the intensive care unit (ICU) of a 400-bed cancer reference center. Simple control measures including contact isolation of colonized patients and reinforcement of handwashing practices among personnel were followed by a sharp decrease in the rate of infection and colonization. An epidemiological investigation showed that a single serophage variant MRSA strain was involved; peak incidence of infection was 17 per 100 ICU patient discharges; the index case was identified as a patient admitted from another hospital and the epidemic strain was then transmitted from patient-to-patient in the ICU; risk factors for acquiring infection were length of prior hospitalization, invasive procedures and number of antibiotic treatments; dissemination of the strain to other wards was only anecdotal. These results stress the effectiveness of simple measures to control outbreaks of MRSA nosocomial infections even in immunocompromised cancer patients.

Cancer Care Facilities

Instability of characteristics amongst coagulase-negative staphylococci causing endocarditis.

Variation in typing of clinically significant isolates of coagulase-negative staphylococci (CNS) was determined by five typing methods with 143 isolates obtained from 19 patients over periods from 2 days to 1 year. In only one case did all isolates give exactly the same typing pattern by all five tests. No single method, or simple combination, provided a ready means of confirming the relatedness of separate isolates. The most frequently useful tests were antibiotic susceptibility and extrachromosomal DNA banding patterns. However, the results of biotyping, serotyping and phage typing were also helpful in showing the relationship between different isolates from a given patient. In most cases a core pattern varying by the gain or loss of a small number of features, characterised a given patient's isolates. In two causes, apparently radical changes in the infecting organism were observed, and confirmed by restriction endonuclease analysis. Care should be taken when successive isolates of CNS show distinct typing differences in deciding their clinical relevance.

Anti-Bacterial Agents

Should we inhibit gastric acid secretion before cardiac surgery?

Stress can decrease intragastric pH and cause erosion of gastric mucosa. Because cardiac surgery and cardiopulmonary bypass represent a major stress, the effects on intragastric pH of an H2-receptor antagonist, ranitidine, and an M1-muscarinic antagonist, pirenzepine, were evaluated. Intragastric pH was measured throughout elective cardiac surgery in 60 patients by a digital pH-meter during fentanyl-diazepam-nitrous oxide (50%) anesthesia. The gastric content was sampled at closure of the chest for bacterial count. Oral preoperative medication given randomly included (n = 20 in each group) 0.3 mg/kg diazepam 1 h before induction (group 1); diazepam plus ranitidine (150 mg) 1 h before induction (group 2); and diazepam plus pirenzepine (50 mg) on the evening before surgery and 1 h before induction of anesthesia (group 3). At induction intragastric pH was higher in group 2 (mean +/- SD = 7.42 +/- 1.07) than in group 1 (5.28 +/- 2.14) (P less than 0.01) but was not significantly different in group 3 (5.78 +/- 1.89) than in group 1. In no group did intragastric pH change significantly during surgery. Gastric juice was sterile in 92% of group 1, in 25% of group 2, and in 71% of group 3 patients (P less than 0.01). Postoperatively no gastrointestinal complications occurred, but there was a trend toward more patients developing nosocomial pneumonias in groups 2 and 3 (15%) than in group 1 (0%) (P = 0.06). Intraoperative intragastric pH is relatively high after diazepam premedication, thus the preoperative addition of ranitidine or pirenzepine would not be necessary and may possibly be hazardous.

Adult

[Frequency and distribution of beta-lactamases in 1792 strains of Klebsiella pneumoniae in France between 1985 and 1988].

In october 1985, 1987 and 1988, all the clinical isolates of K. pneumoniae (respectively 530, 654, 590 strains) were collected in 20 hospitals. The beta-lactamases were identified by analytical isoelectrofocusing and by substrate and inhibition profiles. 76 to 81% of the strains produced only one beta-lactamase: SHV-1 type, pI 7.7 (61 to 65%) or PI 7.1 (14%). The TEM-1 betalactamase (pI 5.4) was produced in 1985 by 21% of the strains, 9% in 1987, and 11% in 1988: TEM-2, pI 5.6 by 2% in 1985-87-88. The extended broad spectrum beta-lactamases, able to hydrolyse amino-thiazol-oximino-beta-lactam antibiotics, TEM or SHV type enzymes (SHV-2, pI 7.7, SHV-3, pI 7.1; SHV-4/CAZ-5, pI 7.8; SHV-5/CAZ-4 pI 8.2; CTX-1/TEM-3, pI 6.3) were also detected: 0.75% of the strain (3 strains) in 1985, 8.4% (55 strains) in 1987, 11% (65 strains) in 1988. These extended broad spectrum beta-lactamases were found in 2 hospitals in 1985, 10 in 1987 and 9 in 1988.

France

Clinical isolates of Staphylococcus lugdunensis and S. schleiferi: bacteriological characteristics and susceptibility to antimicrobial agents.

The bacteriological characteristics and susceptibility to antimicrobial agents of 108 clinical isolates of Staphylococcus lugdunensis and Staphylococcus schleiferi are described. Fifty out of 108 isolates were considered to be responsible for 16 documented infections, including some severe infections (endocarditis, bacteraemia, osteitis). A number of bacteriological characteristics enabled the identification of these species in the clinical microbiology laboratory: the absence of coagulase and protein A, and the presence of a fibrinogen affinity factor and thermonuclease along with other biochemical characteristics (ornithine and arginine decarboxylases, carbohydrate acidification, novobiocin susceptibility) differentiated these new species from other staphylococci; however, they did not possess virulence markers such as toxins or haemagglutinin, but were haemolytic. In this series, almost all isolates were susceptible to 22 antibiotics and 4 antiseptics representative of the main groups of antimicrobial agents. More information is needed on the ecology and epidemiology of these new opportunistic pathogens.

Adult

Hospital dispersion of Staphylococcus epidermidis isolates resistant to a fluoroquinolone, pefloxacin.

Since 1985, nosocomial infections have been frequently treated with a new fluoroquinolone, pefloxacin, at the Cardiological and Neurological Hospital in Lyon. From 1986 to 1988, the incidence of resistance of clinical Staphylococcus epidermidis strains to pefloxacin increased from 31 to 57%. Dispersion of these resistant strains in the hospital environment was recognized when they were detected on 22% of staff members' fingers (139 samples were investigated) and in 28% of the environmental samples (180 were investigated). There was an association between carriage rate and work place. Most of the pefloxacin-resistant S. epidermidis were resistant to oxacillin, gentamicin, erythromycin, cotrimoxazole and fosfomycin. Intensive use of pefloxacin selected multiresistant S. epidermidis which became ubiquitous in the hospital environment.

Carrier State

Susceptibilities to ceftriaxone of streptococcal strains associated with infective endocarditis.

We determined the bactericidal activity of ceftriaxone on 20 streptococci isolated from patients with infective endocarditis and that of penicillin G on 5 strains. The MICs of ceftriaxone were less than or equal to 2 micrograms/ml and the MBCs were low for 5 nontolerant strains (less than or equal to 2 micrograms/ml) and high for 15 tolerant strains (greater than or equal to 16 micrograms/ml). The maximal reduction of the viable bacterial counts after 24 h of exposure to antibiotic was achieved for a concentration of ceftriaxone of 4, 32 and 256 micrograms/ml, respectively for 5, 10 and 19 strains. The activity of penicillin G was similar.

Ceftriaxone

[Sensitivity of Pseudomonas aeruginosa and Klebsiella spp. to ceftazidime. Current status in France].

Ceftazidime was tested against 2,224 strains of Pseudomonas aeruginosa obtained from 17 hospitals in April, May and June, 1986 and against 607 strains of Klebsiella pneumoniae and 234 strains of K. oxytoca obtained from 16 hospitals in October, 1987. The MIC's of ceftazidime against P. aeruginosa were distributed normally, with an MIC50 of 2 mg/l and an MIC90 of 4 mg/l. Depending on critical concentrations, 80 per cent of strains were sensitive, 11.4 per cent were of intermediate sensitivity and 0.54 per cent were resistant. There were few differences in results between hospitals. Ninety-two per cent of resistant strains and 45 per cent of intermediate strains (as opposed to 6 per cent of all strains) produced a high-level constitutive cephalosporinase with little variations between centres. The MIC's of ceftazidime against K. pneumoniae and K. oxytoca had a bimodal distribution: 91 per cent of strains were sensitive to 0.25 mg/l, 6 per cent of strains showed intermediate sensitivity and 3 per cent were resistant. All intermediate and resistant strains produced a very broad spectrum beta-lactamase which hydrolyzed some of the third generation cephalosporins: K. pneumoniae 36 CTX-1, 5 SHV-2, and 14 strains producing a recently identified beta-lactamase "CAZ-5/SHV-4"; K. oxytoca 3 CTX-1. These strains were isolated in 10 of the 16 hospitals which took part in the 1987 study. Comparison of these results with those of studies performed in 1984 and 1985 showed a moderate increase in the number of intermediate sensitivity strains of P. aeruginosa and the occasional occurrence, of the epidemic type, in some hospitals of Klebsiella spp. producing very broad spectrum beta-lactamases which were rare in 1985.

Ceftazidime

Characterization of clinically significant isolates of Staphylococcus epidermidis from patients with endocarditis.

Biotyping, slime production, bacteriophage typing, serotyping, antibiograms, and plasmid profiles were used to characterize 19 Staphylococcus epidermidis strains isolated from 12 patients with prosthetic valve endocarditis and from 7 patients with native valve endocarditis. With the API Staph battery, 12 different biocodes with, at the most, three differences were obtained. Slime production was found for 10 strains (53%). Agglutinogens investigated by agglutination with two specific sera were found for 12 strains (63.1%). Three strains were phage typable (15.2%). Against a panel of nine antimicrobial agents, 15 different profiles were found. Multiply antibiotic-resistant strains were isolated from patients with prosthetic valve endocarditis when disease onset occurred less than 18 months after heart surgery and from patients with native valve endocarditis who received antibiotics immediately prior to their illness. All of the strains were available for plasmid analysis, and all the DNA profiles were distinct. On gels run in Tris-borate buffer, 73.7% of the strains had large plasmids of more than 30 megadaltons. A small plasmid of 2.8 megadaltons was found in multiply resistant strains and in strains resistant only to tetracyclines. None of the isolates appeared to be the same strain, and the bacteriological differences between the strains were confirmed mainly by the antibiotic susceptibility profile and the plasmid pattern analysis. These bacteriological results were in agreement with the clinical data.

Bacteriophage Typing